Anti-Hsp47 antibody [EPR4217]

6 product images

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp47 antibody [EPR4217] (ab109117)

  ab109117 staining Hsp47 in Human colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed and paraffin-embedded, antigen retrieval was by heat mediation in Tris/EDTA buffer pH9. Samples were incubated with primary antibody (1/300). An undiluted HRP-conjugated mouse anti-rabbit IgG was used as the secondary antibody. Tissue counterstained with Hematoxylin.

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  Western blot - Anti-Hsp47 antibody [EPR4217] (ab109117)

  All lanes: Western blot - Anti-Hsp47 antibody [EPR4217] (ab109117) at 1/2000 dilution

  Lane 1: A431 Cell Lysate at 20 µg

  Lane 2: NIH/3T3 Cell Lysate at 20 µg

  Secondary

  All lanes: Goat Anti-Rabbit IgG, (H+L), HRP- conjugated at 1/1000 dilution

  Predicted band size: 46 kDa

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  Western blot - Anti-Hsp47 antibody [EPR4217] (ab109117)

  All lanes: Western blot - Anti-Hsp47 antibody [EPR4217] (ab109117) at 1/1000 dilution

  Lane 1: HeLa cell lysate at 10 µg

  Lane 2: A431 cell lysate at 10 µg

  Lane 3: NIH/3T3 cell lysate at 10 µg

  Predicted band size: 46 kDa

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Hsp47 antibody [EPR4217] (ab109117)

  ab109117, unpurified, at 1/100 dilution, staining Hsp47 in Human breast tissue by Immunohistochemistry.
  

  Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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  Western blot - Anti-Hsp47 antibody [EPR4217] (ab109117)

  Lane 1: Wild-type HAP1 whole cell lysate (20 μg)
  Lane 2: Hsp47 knockout HAP1 whole cell lysate (20 μg)
  Lane 3: HeLa whole cell lysate (20 μg)
  Lane 4: HepG2 whole cell lysate (20 μg)
  
  Lanes 1 - 4: Merged signal (red and green). Green - ab109117 observed at 46 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.
  

  ab109117 was shown to recognize Hsp47 in wild-type HAP1 cells as signal was lost at the expected MW in Hsp47 knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and Hsp47 knockout samples were subjected to SDS-PAGE. ab109117 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-Hsp47 antibody [EPR4217] (ab109117)

  Predicted band size: 46 kDa

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  Image collected and cropped by CiteAb under a CC-BY license from the publication

  Western blot - Anti-Hsp47 antibody [EPR4217] (ab109117)

  Hsp47 western blot using anti-Hsp47 antibody [EPR4217] ab109117. Publication image and figure legend from Tian, S., Peng, P., et al., 2020, Aging (Albany NY), PubMed 32091407.

  
  

  ab109117 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab109117 please see the product overview.

  SERPINH1 regulates EMT markers and Wnt/β-catenin signaling pathway in GC. (A) Western blot analysis shows reduced N-cadherin and increased E-cadherin expression in SERPINH1-silenced SGC-7901 cells compared with controls. Conversely, SERPINH1-overexpressing MGC-803 cells show increased N-cadherin and reduced E-cadherin expression compared with the controls. (B) Western blot analysis shows increased levels of β-catenin, Wnt2, GSK-3β, p-GSK-3β, NF-κB p65, Snail1, Slug, and TWIST in the SERPINH1-overexpressing MGC-803 cells compared with the controls, whereas SERPINH1-silenced SGC-7901 cells show reduced levels of β-catenin, Wnt2, GSK-3β, p-GSK-3β, NF-κB p65, Snail1, Slug, and TWIST compared with the controls. (C) Immunofluorescence staining of E-cadherin, N-cadherin and SERPINH1 proteins in the control and SERPINH1-silenced SGC-7901 cells, as well as, control and SERPINH1-overexpressing MGC-803 cells. (D, E) Gene expression analysis of the TCGA-STAD dataset shows (D) negative correlation of CDH1 (r=-0.12, P=0.019) or E-cadherin mRNA levels and (E) positive association of CDH2 (r=0.40, P<0.0001) or N-cadherin mRNA levels with the SERPINH1 mRNA levels.